Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging

Mary P. Hall, Carolyn C. Woodroofe, Monika G. Wood, Ivo Que, Moniek Root, Yanto Ridwan, Ce Shi, Thomas A. Kirkland, Lance P. Encell, Keith V. Wood, Clemens Löwik and Laura Mezzanotte ()
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Mary P. Hall: Promega Corporation
Carolyn C. Woodroofe: Promega Corporation
Monika G. Wood: Promega Corporation
Ivo Que: Leiden University Medical Center
Moniek Root: Erasmus University Medical Center
Yanto Ridwan: Erasmus University Medical Center
Ce Shi: Promega Biosciences Incorporated
Thomas A. Kirkland: Promega Biosciences Incorporated
Lance P. Encell: Promega Corporation
Keith V. Wood: Promega Corporation
Clemens Löwik: Erasmus University Medical Center
Laura Mezzanotte: Erasmus University Medical Center

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: Abstract The sensitivity of bioluminescence imaging in animals is primarily dependent on the amount of photons emitted by the luciferase enzyme at wavelengths greater than 620 nm where tissue penetration is high. This area of work has been dominated by firefly luciferase and its substrate, D-luciferin, due to the system’s peak emission (~ 600 nm), high signal to noise ratio, and generally favorable biodistribution of D-luciferin in mice. Here we report on the development of a codon optimized mutant of click beetle red luciferase that produces substantially more light output than firefly luciferase when the two enzymes are compared in transplanted cells within the skin of black fur mice or in deep brain. The mutant enzyme utilizes two new naphthyl-luciferin substrates to produce near infrared emission (730 nm and 743 nm). The stable luminescence signal and near infrared emission enable unprecedented sensitivity and accuracy for performing deep tissue multispectral tomography in mice.

Date: 2018
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DOI: 10.1038/s41467-017-02542-9

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